Establishing a metastasis-related diagnosis and prognosis model for lung adenocarcinoma through CRISPR library and TCGA database.

PURPOSE: Existing biomarkers for diagnosing and predicting metastasis of lung adenocarcinoma (LUAD) may not meet the demands of clinical practice. Risk prediction models with multiple markers may provide better prognostic factors for accurate diagnosis and prediction of metastatic LUAD. METHODS: An animal model of LUAD metastasis was constructed using CRISPR technology, and genes related to LUAD metastasis were screened by mRNA sequencing of normal and metastatic tissues. The immune characteristics of different subtypes were analyzed, and differentially expressed genes were subjected to survival and Cox regression analyses to identify the specific genes involved in metastasis for constructing a prediction model. The biological function of RFLNA was verified by analyzing CCK-8, migration, invasion, and apoptosis in LUAD cell lines. RESULTS: We identified 108 differentially expressed genes related to metastasis and classified LUAD samples into two subtypes according to gene expression. Subsequently, a prediction model composed of eight metastasis-related genes (RHOBTB2, KIAA1524, CENPW, DEPDC1, RFLNA, COL7A1, MMP12, and HOXB9) was constructed. The areas under the curves of the logistic regression and neural network were 0.946 and 0.856, respectively. The model effectively classified patients into low- and high-risk groups. The low-risk group had a better prognosis in both the training and test cohorts, indicating that the prediction model had good diagnostic and predictive power. Upregulation of RFLNA successfully promoted cell proliferation, migration, invasion, and attenuated apoptosis, suggesting that RFLNA plays a role in promoting LUAD development and metastasis. CONCLUSION: The model has important diagnostic and prognostic value for metastatic LUAD and may be useful in clinical applications.

A nomogram prediction of pressure injury in critical ill patients: A retrospective cohort study.

Pressure injury (PI) is still a significant public health problem to be solved. Accurate prediction can lead to timely prophylaxis and therapy. However, the currently used Braden score shows insufficient predictive validity. We aimed to develop a nomogram to predict PI development in critically ill patients. We extracted data from Medical Information Mart for Intensive Care-IV v1.0. Variable selection was based on univariate logistic regression and all-subset regression. The area under the receiver operating characteristic curve (AUC) was used to assess the performance of the nomogram and Braden score. Decision curve analysis (DCA) was performed to identify and compare the clinical usefulness between the nomogram model and Braden score. We have developed a novel and practical nomogram that accurately predicts pressure ulcers. The AUC of the new model was better than that of the Braden score (P < .001). DCA showed that the nomogram model had a better net benefit than the Braden score at any given threshold. This finding needs to be confirmed by external validation as well as multicentre prospective studies.

Value of early critical care transthoracic echocardiography for patients undergoing mechanical ventilation: a retrospective study.

OBJECTIVES: To evaluate whether early intensive care transthoracic echocardiography (TTE) can improve the prognosis of patients with mechanical ventilation (MV). DESIGN: A retrospective cohort study. SETTING: Patients undergoing MV for more than 48 hours, based on the Medical Information Mart for Intensive Care III (MIMIC-III) database and the eICU Collaborative Research Database (eICU-CRD), were selected. PARTICIPANTS: 2931 and 6236 patients were recruited from the MIMIC-III database and the eICU database, respectively. PRIMARY AND SECONDARY OUTCOME MEASURES: The primary outcome was in-hospital mortality. Secondary outcomes were 30-day mortality from the date of ICU admission, days free of MV and vasopressors 30 days after ICU admission, use of vasoactive drugs, total intravenous fluid and ventilator settings during the first day of MV. RESULTS: We used propensity score matching to analyse the association between early TTE and in-hospital mortality and sensitivity analysis, including the inverse probability weighting model and covariate balancing propensity score model, to ensure the robustness of our findings. The adjusted OR showed a favourable effect between the early TTE group and in-hospital mortality (MIMIC: OR 0.78; 95% CI 0.65 to 0.94, p=0.01; eICU-CRD: OR 0.76; 95% CI 0.67 to 0.86, p<0.01). Early TTE was also associated with 30-day mortality in the MIMIC database (OR 0.71, 95% CI 0.57 to 0.88, p=0.001). Furthermore, those who had early TTE had both more ventilation-free days (only in eICU-CRD: 23.48 vs 24.57, p<0.01) and more vasopressor-free days (MIMIC: 18.22 vs 20.64, p=0.005; eICU-CRD: 27.37 vs 28.59, p<0.001) than the control group (TTE applied outside of the early TTE and no TTE at all). CONCLUSIONS: Early application of critical care TTE during MV is beneficial for improving in-hospital mortality. Further investigation with prospectively collected data is required to validate this relationship.

Phospho-Tyr705 of STAT3 is a therapeutic target for sepsis through regulating inflammation and coagulation.

BACKGROUND: Sepsis is an infection-induced aggressive and life-threatening organ dysfunction with high morbidity and mortality worldwide. Infection-associated inflammation and coagulation promote the progression of adverse outcomes in sepsis. Here, we report that phospho-Tyr705 of STAT3 (pY-STAT3), not total STAT3, contributes to systemic inflammation and coagulopathy in sepsis. METHODS: Cecal ligation and puncture (CLP)-induced septic mice were treated with BP-1-102, Napabucasin, or vehicle control respectively and then assessed for systemic inflammation, coagulation response, lung function and survival. Human pulmonary microvascular endothelial cells (HPMECs) and Raw264.7 cells were exposed to lipopolysaccharide (LPS) with pharmacological or genetic inhibition of pY-STAT3. Cells were assessed for inflammatory and coagulant factor expression, cell function and signaling. RESULTS: Pharmacological inhibition of pY-STAT3 expression by BP-1-102 reduced the proinflammatory factors, suppressed coagulation activation, attenuated lung injury, alleviated vascular leakage and improved the survival rate in septic mice. Pharmacological or genetic inhibition of pY-STAT3 diminished LPS-induced cytokine production in macrophages and protected pulmonary endothelial cells via the IL-6/JAK2/STAT3, NF-κB and MAPK signaling pathways. Moreover, the increase in procoagulant indicators induced by sepsis such as tissue factor (TF), the thrombin-antithrombin complex (TAT) and D-Dimer were down-regulated by pY-STAT3 inhibition. CONCLUSIONS: Our results revealed a therapeutic role of pY-STAT3 in modulating the inflammatory response and defective coagulation during sepsis. Video Abstract.

[Study on Serological Blood Group Conversion in BM Empty Phase of ABO-incompatible Allogeneic Stem Cell Transplantation].

OBJECTIVE: To explore the regularity of serological conversion of blood group in BM empty phase of ABO-incompatible allogeneic stem cell transplantation so as to provide the basis for selecting the blood components in blood transfusion. METHODS: Before hematpoietic stem cell transplantation (HSCT), the ABO and RhD blood groups of recipients and donors were identified by salt medium tube method and microcolumn gel method; after transplantation the changes of antigen intensity and antibody titer of ABO blood group in patients were periodically detected. RESULTS: After blood group shift of 33 patients received ABO-incompatible allo-HSCT, the consistent rate of positive and regative types in major ABO incompatible group was 100%; the consistent rate of positive and negative types in minor ABO-incompatible group was 33%, no-consistent rate was 66.7%; the consistent rate of positive and negative types in bidirextional ABO incompatible group was 20%, the no-consistent rate was 80%. CONCLUSION: After ABO-incompatible allo-HSCT, blood group antgen of patients shifts to the blood group of donors, there is a significant difference in the serological indicators between the minor and bidireetional ABO-incompatible patients and normal people.

miR-326 targets antiapoptotic Bcl-xL and mediates apoptosis in human platelets.

Platelets play crucial roles in hemostasis, thrombosis, wound healing, inflammation, angiogenesis, and tumor metastases. Because they are anucleated blood cells, platelets lack nuclear DNA, but they do contain mitochondrial DNA, which plays a key role in regulating apoptosis. Recent evidence has suggested that miRNAs are also involved in regulating gene expression and apoptosis in platelets. Our previous study showed that the expression of miR-326 increased visibly when apheresis platelets were stored in vitro. The antiapoptotic Bcl-2 family regulator Bcl-xL has been identified as a putative target of miR-326. In the present study, dual reporter luciferase assays were used to characterize the function of miR-326 in the regulation of the apoptosis of platelet cells. These assays demonstrated that miR-326 bound to the 3'-translated region of Bcl-xL. To directly assess the functional effects of miR-326 expression, levels of Bcl-xL and the apoptotic status of stored apheresis platelets were measured after transfection of miR-326 mimic or inhibitor. Results indicated that miR-326 inhibited Bcl-xL expression and induced apoptosis in stored platelets. Additionally, miR-326 inhibited Bcl-2 protein expression and enhanced Bak expression, possibly through an indirect mechanism, though there was no effect on the expression of Bax. The effect of miR-326 appeared to be limited to apoptosis, with no significant effect on platelet activation. These results provide new insight into the molecular mechanisms affecting differential platelet gene regulation, which may increase understanding of the role of platelet apoptosis in multiple diseases.